LOCUS       sequence-436893-        8561 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express HACE Editor as a fusion of AID cytidine deaminase and BLM 
            helicase..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8561)
  AUTHORS   Chen XD, Chen Z, Wythes G, Zhang Y, Orr BC, Sun G, Chao YK, Navarro 
            Torres A, Thao K, Vallurupalli M, Sun J, Borji M, Tkacik E, Chen H, 
            Bernstein BE, Chen F
  TITLE     Helicase-assisted continuous editing for programmable mutagenesis of
            endogenous genomes.
  JOURNAL   Science. 2024 Oct 11;386(6718):eadn5876. doi: 
            10.1126/science.adn5876. Epub 2024 Oct 11.
  PUBMED    39388570
REFERENCE   2  (bases 1 to 8561)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Science."; 
            date: "2024-10-11"; volume: "386(6718):eadn5876. doi"; pages: " 
            10.1126/science.adn5876. Epub 2024 Oct 11"
COMMENT     Sequence Label: pCMV-AID-BLM-UGI
FEATURES             Location/Qualifiers
     source          1..8561
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        132..335
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     286..306
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      384..393
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5286..5534
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             5547..5567
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5576..5593
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5616..5633)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5622..5846
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5917..5933)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5917..5933)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5930..5952)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5941..5957
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5965..5971,5972..5989,5990..5995))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6010..6031
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6148..6165)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6319..6907)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6399..6418)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7078..7869,7870..7938))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7701..7720
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7939..8043)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8122..8141)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        join(8313..8561,1..131)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
       61 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      121 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      181 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      241 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      301 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
      361 agagatccgc ggccgcgaga gccgccacca tggacagcct cttgatgaac cggagggagt
      421 ttctttacca attcaaaaat gtccgctggg ctaagggtcg gcgtgagacc tacctgtgct
      481 acgtagtgaa gaggcgtgac agtgctacat ccttttcact ggactttggt tatcttcgca
      541 ataagaacgg ctgccacgtg gaattgctct tcctccgcta catctcggac tgggacctag
      601 accctggccg ctgctaccgc gtcacctggt tcatctcctg gagcccctgc tacgactgtg
      661 cccgacatgt ggccgacttt ctgcgaggga accccaacct cagtctgagg atcttcaccg
      721 cgcgcctcta cttctgtgag gaccgcaagg ctgagcccga ggggctgcgg cggctgcacc
      781 gcgccggggt gcaaatagcc atcatgacct tcaaagatta tttttactgc tggaatactt
      841 ttgtagaaaa ccacggaaga actttcaaag cctgggaagg gctgcatgaa aattcagttc
      901 gtctctccag acagcttcgg cgcatccttt tgcccctgta tgaggttgat gacttacgag
      961 acgcatttcg tactagcggc agcgagactc ccgggacctc agagtccgcc acacccgaaa
     1021 gtatggctgc tgttcctcaa aataatctac aggagcaact agaacgtcac tcagccagaa
     1081 cacttaataa taaattaagt ctttcaaaac caaaattttc aggtttcact tttaaaaaga
     1141 aaacatcttc agataacaat gtatctgtaa ctaatgtgtc agtagcaaaa acacctgtat
     1201 taagaaataa agatgttaat gttaccgaag acttttcctt cagtgaacct ctacccaaca
     1261 ccacaaatca gcaaagggtc aaggacttct ttaaaaatgc tccagcagga caggaaacac
     1321 agagaggtgg atcaaaatca ttattgccag atttcttgca gactccgaag gaagttgtat
     1381 gcactaccca aaacacacca actgtaaaga aatcccggga tactgctctc aagaaattag
     1441 aatttagttc ttcaccagat tctttaagta ccatcaatga ttgggatgat atggatgact
     1501 ttgatacttc tgagacttca aaatcatttg ttacaccacc ccaaagtcac tttgtaagag
     1561 taagcactgc tcagaaatca aaaaagggta agagaaactt ttttaaagca cagctttata
     1621 caacaaacac agtaaagact gacttgcctc caccctcctc tgaaagcgag caaatagatt
     1681 tgactgagga acagaaggat gactcagaat ggttaagcag cgatgtgatt tgcatcgatg
     1741 atggccccat tgctgaagtg catataaatg aagatgctca ggaaagtgac tctctgaaaa
     1801 ctcatttgga agatgaaaga gataatagcg aaaagaagaa gaatttggaa gaagctgaat
     1861 tacattcaac tgagaaagtt ccatgtattg aatttgatga tgatgattat gatacggatt
     1921 ttgttccacc ttctccagaa gaaattattt ctgcttcttc ttcctcttca aaatgcctta
     1981 gtacgttaaa ggaccttgac acctctgaca gaaaagagga tgttcttagc acatcaaaag
     2041 atcttttgtc aaaacctgag aaaatgagta tgcaggagct gaatccagaa accagcacag
     2101 actgtgacgc tagacagata agtttacagc agcagcttat tcatgtgatg gagcacatct
     2161 gtaaattaat tgatactatt cctgatgata aactgaaact tttggattgt gggaacgaac
     2221 tgcttcagca gcggaacata agaaggaaac ttctaacgga agtagatttt aataaaagtg
     2281 atgccagtct tcttggctca ttgtggagat acaggcctga ttcacttgat ggccctatgg
     2341 agggtgattc ctgccctaca gggaattcta tgaaggagtt aaatttttca caccttccct
     2401 caaattctgt ttctcctggg gactgtttac tgactaccac cctaggaaag acaggattct
     2461 ctgccaccag gaagaatctt tttgaaaggc ctttattcaa tacccattta cagaagtcct
     2521 ttgtaagtag caactgggct gaaacaccaa gactaggaaa aaaaaatgaa agctcttatt
     2581 tcccaggaaa tgttctcaca agcactgctg tgaaagatca gaataaacat actgcttcaa
     2641 taaatgactt agaaagagaa acccaacctt cctatgatat tgataatttt gacatagatg
     2701 actttgatga tgatgatgac tgggaagaca taatgcataa tttagcagcc agcaaatctt
     2761 ccacagctgc ctatcaaccc atcaaggaag gtcggccaat taaatcagta tcagaaagac
     2821 tttcctcagc caagacagac tgtcttccag tgtcatctac tgctcaaaat ataaacttct
     2881 cagagtcaat tcagaattat actgacaagt cagcacaaaa tttagcatcc agaaatctga
     2941 aacatgagcg tttccaaagt cttagttttc ctcatacaaa ggaaatgatg aagatttttc
     3001 ataaaaaatt tggcctgcat aattttagaa ctaatcagct agaggcgatc aatgctgcac
     3061 tgcttggtga agactgtttt atcctgatgc cgactggagg tggtaagagt ttgtgttacc
     3121 agctccctgc ctgtgtttct cctggggtca ctgttgtcat ttctcccttg agatcactta
     3181 tcgtagatca agtccaaaag ctgacttcct tggatattcc agctacatat ctgacaggtg
     3241 ataagactga ctcagaagct acaaatattt acctccagtt atcaaaaaaa gacccaatca
     3301 taaaactcct atatgtcact ccagaaaaga tctgtgcaag taacagactc atttctactc
     3361 tggagaatct ctatgagagg aagctcttgg cacgttttgt tattgatgaa gcacattgtg
     3421 tcagtcagtg gggacatgat tttcgtcaag attacaaaag aatgaatatg cttcgccaga
     3481 agtttccttc tgttccggtg atggctctta cggccacagc taatcccagg gtacagaagg
     3541 acatcctgac tcagctgaag attctcagac ctcaggtgtt tagcatgagc tttaacagac
     3601 ataatctgaa atactatgta ttaccgaaaa agcctaaaaa ggtggcattt gattgcctag
     3661 aatggatcag aaagcaccac ccatatgatt cagggataat ttactgcctc tccaggcgag
     3721 aatgtgacac catggctgac acgttacaga gagatgggct cgctgctctt gcttaccatg
     3781 ctggcctcag tgattctgcc agagatgaag tgcagcagaa gtggattaat caggatggct
     3841 gtcaggttat ctgtgctaca attgcatttg gaatggggat tgacaaaccg gacgtgcgat
     3901 ttgtgattca tgcatctctc cctaaatctg tggagggtta ctaccaagaa tctggcagag
     3961 ctggaagaga tggggaaata tctcactgcc tgcttttcta tacctatcat gatgtgacca
     4021 gactgaaaag acttataatg atggaaaaag atggaaacca tcatacaaga gaaactcact
     4081 tcaataattt gtatagcatg gtacattact gtgaaaatat aacagaatgc aggagaatac
     4141 agcttttggc ctactttggt gaaaatggat ttaatcctga tttttgtaag aaacacccag
     4201 atgtttcttg tgataattgc tgtaaaacaa aggattataa aacaagagat gtgactgacg
     4261 atgtgaaaag tattgtaaga tttgttcaag aacatagttc atcacaagga atgagaaata
     4321 taaaacatgt aggtccttct ggaagattta ctatgaatat gctggtcgac attttcttgg
     4381 ggagtaagag tgcaaaaatc cagtcaggta tatttggaaa aggatctgct tattcacgac
     4441 acaatgccga aagacttttt aaaaagctga tacttgacaa gattttggat gaagacttat
     4501 atatcaatgc caatgaccag gcgatcgctt atgtgatgct cggaaataaa gcacaaactg
     4561 tactaaatgg caatttaaag gtagacttta tggaaacaga aaattccagc agtgtgaaaa
     4621 aacaaaaagc gttagtagca aaagtgtctc agagggaaga gatggttaaa aaatgtcttg
     4681 gagaacttac agaagtctgc aaatctctgg ggaaagtttt tggtgtccat tacttcaata
     4741 tttttaatac cgtcactctc aagaagcttg cagaatcttt atcttctgat cctgaggttt
     4801 tgcttcaaat tgatggtgtt actgaagaca aactggaaaa atatggtgcg gaagtgattt
     4861 cagtattaca gaaatactct gaatggacat cgccagctga agacagttcc ccagggataa
     4921 gcctgtccag cagcagaggc cccggaagaa gtgccgctga ggagcttgac gaggaaatac
     4981 ccgtatcttc ccactacttt gcaagtaaaa ccagaaatga aaggaagagg aaaaagatgc
     5041 cagcctccca aaggtctaag aggagaaaaa ctgcttccag tggttccaag gcaaaggggg
     5101 ggtctgccac atgtagaaag atatcttcca aaacgaaatc ctccagcatc attggatcca
     5161 gttcagcctc acatacttct caagcgacat caggagccaa tagcaaattg gggattatgg
     5221 ctccaccgaa gcctataaat agaccgtttc ttaagccttc atatgcattc tcatctggtg
     5281 gttctactaa tctgtcagat attattgaaa aggagaccgg taagcaactg gttatccagg
     5341 aatccatcct catgctccca gaggaggtgg aagaagtcat tgggaacaag ccggaaagcg
     5401 atatactcgt gcacaccgcc tacgacgaga gcaccgacga gaatgtcatg cttctgacta
     5461 gcgacgcccc tgaatacaag ccttgggctc tggtcataca ggatagcaac ggtgagaaca
     5521 agattaagat gctctctggt ggttctccca agaagaagag gaaagtctaa ccggtcatca
     5581 tcaccatcac cattgagttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca
     5641 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     5701 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     5761 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     5821 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctcgat
     5881 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     5941 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagccta
     6001 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     6061 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     6121 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     6181 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     6241 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     6301 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     6361 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     6421 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     6481 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     6541 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     6601 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     6661 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     6721 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     6781 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     6841 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     6901 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     6961 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     7021 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     7081 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     7141 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     7201 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     7261 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     7321 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     7381 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     7441 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     7501 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     7561 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     7621 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     7681 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     7741 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     7801 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     7861 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     7921 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     7981 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     8041 gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc gatctcccga
     8101 tcccctaggg tcgactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct
     8161 gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac
     8221 aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct
     8281 gcttcgcgat gtacgggcca gatatacgcg ttgacattga ttattgacta gttattaata
     8341 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
     8401 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
     8461 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
     8521 tttacggtaa actgcccact tggcagtaca tcaagtgtat c
//
